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Updated: Mar 22, 2026
Detection and Isolation of Viable Mouse IL-17-Secreting T Cells
Published on: December 18, 2008
[Preparation and identification of the polyclonal antibody against mouse IL-23p19]
Yingli Men1, Xiaolong Wang1, Cong Ding1
1Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
Objective:
To prepare the polyclonal antibody against mouse interleukine-23 p19 (IL-23p19).
Methods:
The murine full-length IL-23p19 gene was subcloned into pET16 expression vector to construct the recombinant plasmid pET-16b-IL-23p19. The plasmid was transformed into E. coli BL 21 (DE3) and IL-23p19 protein expression was induced by IPTG and identified with SDS-PAGE analysis and Western blotting. After purified by Ni(+) affinity column chromatography, the IL-23p19 protein was used as the antigen to immunize New Zealand rabbit to prepare the antiserum. The polyclonal antibody against the mouse IL-23p19 was isolated from antiserum by affinity chromatography. Antibody titer was detected by ELISA. Antibody specificity was evaluated by Western blotting.
Results:
The pET-16b-IL-23p19 recombinant plasmid was successfully constructed and the IL-23p19 protein was effectively expressed in E. coli BL 21 (DE3). The antibody was successfully prepared by immunizing New Zealand rabbit with the IL-23p19 protein four times. ELISA showed that the titer of the anti-mouse IL-23p19 polyclonal antibody was about 1:256,000. Western blotting confirmed that anti-mouse IL-23p19 polyclonal antibody could specifically recognize the IL-23p19 protein.
Conclusion:
We have successfully prepared the anti-IL-23p19 polyclonal antibody with the high titer and specificity.
